Annexin V-PE Apoptosis Detection Kit

Antigen: Annexin A5 (ANXA5)

Synonyme: PP4, ANX5, ENX2, Anx5, R74653, LC5, MGC93655, ANXA5, MGC126971, anx, anx5, ANX V, anxa5, cb989, wu:fa98f06, wu:fj10f10, MGC89158

Konjugat: PE

Applikation: Durchflusszytometrie (FACS), Fluoreszenzmikroskopie (FM)

Produktnummer: ABIN411992

Produktbeschreibung

Weitere Bezeichnung: Annexin V

Beschreibung: The Annexin V-PE Apoptosis Detection Kit is based on the observation that soon after initiating apoptosis, most cell types translocate the membrane phospholipid phosphatidylserine (PS) from the inner face of the plasma membrane to the cell surface. Once on the cell surface, PS can be easily detected by staining with a fluorescent conjugate of Annexin V, a protein that has a strong natural affinity for PS. The one-step staining procedure takes only 10 minutes. In addition, the assay can be directly performed on live cells, without the need for fixation. The Annexin V-PE Apoptosis Detection Kit contains the bright orange-red PE fluorescent probe that can be easily detected by flow cytometry or fluorescence microscopy.

Anwendungen

Protokoll: A. Incubation of cells with Annexin V-PE 1. Induce apoptosis by desired method. 2. Collect 1-5 x 105 cells by centrifugation. 3. Resuspend cells in 500 µl of 1X Binding Buffer. 4. Add 5 µl of Annexin V-PE. 5. Incubate at room temperature for 5 min in the dark. Proceed to B or C below depending on method of analysis. B. Quantification by Flow Cytometry Analyze Annexin V-PE binding by flow cytometry (Ex = 488 nm, Em = 578 nm) using the phycoerythrin emission signal detector (usually FL2). For analyzing adherent cells, gently trypsinize and wash cells once with serum-containing media before incubation with Annexin V-PE (A.3-5). C. Detection by Fluorescence Microscopy 1. Place the cell suspension from Step A.5 on a glass slide. Cover the cells with a glass coverslip. For analyzing adherent cells, grow cells directly on a coverslip. Following incubation (A.5), invert coverslip on glass slide and visualize cells. The cells can also be washed and fixed in 2% formaldehyde before visualization. (Cells must be incubated with Annexin V-PE before fixation since any cell membrane disruption can cause nonspecific binding of Annexin V to PS on the inner surface of the cell membrane.) 2. Observe the cells under a fluorescence microscope using a rhodamine filter. Cells which have bound Annexin V-PE will show orange-red staining in the plasma membrane.

Lagerung: Store kit at 4 o C. All reagents are stable for one year under proper storage conditions. V. Apoptosis Detection Kits & Reagents • Annexin V Kits & Bulk Reagents • Caspase Assay Kits & Reagents • Mitochondrial Apoptosis Kits & Reagents • Nuclear Apoptosis Kits & Reagents • Apoptosis Inducers and Set • Apoptosis siRNA Vectors Cell Fractionation System • Mitochondria/Cytosol Fractionation Kit • Nuclear/Cytosol Fractionation Kit • Membrane Protein Extraction Kit • Cytosol/Particulate Rapid Separation Kit • Mammalian Cell Extraction Kit • FractionPREP Fractionation System Cell Proliferation & Senescence • Quick Cell Proliferation Assay Kit • Senescence Detection Kit • High Throughput Apoptosis/Cell Viability Assay Kits • LDH-Cytotoxicity Assay Kit • Bioluminescence Cytotoxicity Assay Kit • Live/Dead Cell Staining Kit Cell Damage & Repair • HDAC Fluorometric & Colorimetric Assays & Drug Discovery Kits • HAT Fluorometric & Colorimetric Assays & Drug Discovery Kits • DNA Damage Quantification Kit • Glutathione & Nitric Oxide Fluorometric & Colorimetric Assay Kits Signal Transduction • cAMP & cGMP Assay Kits • Akt & JNK Activity Assay Kits • Beta-Secretase Activity Assay Kit Adipocyte & Lipid Transfer • Recombinant Adiponectin, Survivin, & Leptin • CETP & PLTP Activity Assay & Drug Discovery Kits • Cholesterol Quantification Kit Molecular Biology & Reporter Assays • siRNA Expression Vectors • Cloning Insert Quick Screening Kit • Mitochondrial & Genomic DNA Isolation Kits • 5 Minutes DNA Ligation Kit • 20 Minutes Gel Staining/Destaining Kit • β -Galactosidase Staining Kit & Luciferase Reporter Assay Kit Growth Factors and Cytokine Monoclonal and Polyclonal Antibodies Components K128-25 K128-100 K128-400 25 assays 100 assays 400 assays Annexin V-PE 125 μ l 500 μ l 2 ml 1X Binding Buffer 12.5 ml 50 ml 2 x 100 ml

Beschränkungen: Nur für Forschungszwecke einsetzbar

Publikationen

Publikationen: Zhu, Wang, Wang et al.: "Hepatic deletion of Smad7 in mouse leads to spontaneous liver dysfunction and aggravates alcoholic liver injury." in: PLoS ONE, Vol. 6, Issue 2, pp. e17415, 2011 (PubMed). Weitere Details: Cells were stained for 5 minutes with Annexin V-PE Apoptosis Detection Kit